Cell proliferation analyses are crucial for cell growth and differentiation studies, and are often used to evaluate both compound toxicity and inhibition of tumor cell growth during drug development. Proliferation measurements are typically made based on average DNA content or on cellular metabolism parameters. Assays can report either total or live cell numbers, or measure DNA synthesis in single cells.

Molecular Probes® cell proliferation assays can be used individually or together to monitor cell health and cell division by evaluating DNA synthesis, monitoring mitotic phases, or tracking population doubling.

DNA synthesis measurement assays

Measuring the synthesis of new DNA is a precise way to assay cell proliferation in individual cells or in cell populations. DNA synthesis–based cell proliferation assays measure the rate of new DNA synthesis based on incorporation of modified nucleosides. The Click-iT® Plus EdU cell proliferation assay utilizes the power of click chemistry and the modified nucleoside EdU to provide a superior alternative to BrdU staining for detecting and quantitating newly synthesized DNA.

Measuring cell proliferation with Click-iT® Plus EdU assays also means that cell proliferation measurements can be easily multiplexed with expressed proteins like GFP, protein labels like R-PE, and a wide range of organic fluorophores.

Accuracy—superior to BrdU assays with minimal variation (low CVs)

Simplicity—streamlined five-step protocol

Compatibility—multiplex with GFP, mCherry, APC, PerCP, PE, and other fluorophores

Dye dilution assays for cell proliferation

Dye dilution assays for cell proliferation rely on cell membrane–permeant fluorescent molecules. Upon entry into the cell, the dye will covalently bind to amine groups on proteins, resulting in long-term dye retention within the cell.

Through subsequent cell divisions, each daughter cell receives approximately half the fluorescence of the parent. Analysis of the fluorescence intensities of cell populations by flow cytometry enables determination of the number of generations through which a cell or population has progressed since the label was applied.

CellTrace™ fluorescent stains can be used without affecting morphology or physiology to trace generations in vivo or in vitro.

Long-term signal stability—well-retained in cells for several days post-stain

Noncytotoxic—no known effect on proliferative ability or biology of cells